All living organisms regulate biological function via molecular signaling and metabolism. Currently, the ability to track metabolic transformations deep inside tissue pales when compared to the critical role metabolism plays in living systems. It is our goal to establish technology to map metabolites through metabolic pathways directly in vivo. Specifically, we propose 3D hyperpolarized magnetic resonance imaging of NAD (nicotinamide adenine dinucleotide) and its metabolic transformations in individual organs. The NAD regulates major cellular events, including bioenergetics and signaling pathways such as enzyme regulation, control of gene expression and DNA repair. In fact, NAD homeostasis is strongly affected during disease states such as diabetes, Alzheimer?s or cancer. In addition, current interest has been spiked by findings that show NAD precursor supplementation to improve brain function and expand life span. In mammals, NAD is synthesized from vitamin B3 precursors, most prominently from nicotinamide. The proposed technology will track hyperpolarized signals from nicotinamide and all its downstream metabolites in real time to create 3D maps of NAD pathways and their kinetics. In addition, the proposed technology is cost-efficient and free of adverse effects, essentially using a vitamin shot for 3D molecular imaging. Our team has already demonstrated the physical principles of the methods; the proposed work transforms our basic physics and chemistry advances into a practical, general, and affordable technology which enables unprecedented insights into metabolic regulation, and has a clear translational path for scanning broad patient populations.